SOLUBILIZATION AND CHARACTERIZATION OF MICROSOMAL‐ASSOCIATED PHOSPHATIDYLINOSITOL: CERAMIDE PHOSPHOINOSITOL TRANSFERASE FROM SACCHAROMYCES CEREVISIAE

JESANG KO; SHWUYENG CHEAH; ANTHONY S. FISCHL

doi:10.1111/j.1745-4514.1995.tb00533.x

SOLUBILIZATION AND CHARACTERIZATION OF MICROSOMAL‐ASSOCIATED PHOSPHATIDYLINOSITOL: CERAMIDE PHOSPHOINOSITOL TRANSFERASE FROM SACCHAROMYCES CEREVISIAE

JESANG KO, SHWUYENG CHEAH, ANTHONY S. FISCHL

Research output: Contribution to journal › Article › peer-review

9 Citations (Scopus)

Abstract

The membrane associated enzyme phosphatidylinositol: ceramide phosphoinositol transferase (IPC synthase) catalyzes an essential step in the biosynthesis of yeast inositol‐containing sphingolipids. A variety of solubilization agents were examined for their ability to release IPC synthase activity from yeast membranes. The most effective solubilization agent was Triton X‐100 which released over 90% of the IPC synthase activity. The basic enzymological properties of the solubilized enzyme were determined using a Triton X‐100/phosphatidylinositol/ceramide/enzyme mixed micellar assay system. IPC synthase activity was dependent upon the surface concentrations of phosphatidylinositol and ceramide in a phosphatidylinositol/ceramide/Triton X‐100 mixed micelle. Maximal enzyme activity was measured at 30C and pH 7.0 in the presence of 5 mM Triton X‐100, 1 mM manganese and 5 mM magnesium ions.

Original language	English
Pages (from-to)	253-267
Number of pages	15
Journal	Journal of Food Biochemistry
Volume	19
Issue number	4
DOIs	https://doi.org/10.1111/j.1745-4514.1995.tb00533.x
Publication status	Published - 1995 Aug
Externally published	Yes

ASJC Scopus subject areas

Food Science
Biophysics
Pharmacology
Cell Biology

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title = "SOLUBILIZATION AND CHARACTERIZATION OF MICROSOMAL‐ASSOCIATED PHOSPHATIDYLINOSITOL: CERAMIDE PHOSPHOINOSITOL TRANSFERASE FROM SACCHAROMYCES CEREVISIAE",

abstract = "The membrane associated enzyme phosphatidylinositol: ceramide phosphoinositol transferase (IPC synthase) catalyzes an essential step in the biosynthesis of yeast inositol‐containing sphingolipids. A variety of solubilization agents were examined for their ability to release IPC synthase activity from yeast membranes. The most effective solubilization agent was Triton X‐100 which released over 90% of the IPC synthase activity. The basic enzymological properties of the solubilized enzyme were determined using a Triton X‐100/phosphatidylinositol/ceramide/enzyme mixed micellar assay system. IPC synthase activity was dependent upon the surface concentrations of phosphatidylinositol and ceramide in a phosphatidylinositol/ceramide/Triton X‐100 mixed micelle. Maximal enzyme activity was measured at 30C and pH 7.0 in the presence of 5 mM Triton X‐100, 1 mM manganese and 5 mM magnesium ions.",

author = "JESANG KO and SHWUYENG CHEAH and FISCHL, {ANTHONY S.}",

year = "1995",

month = aug,

doi = "10.1111/j.1745-4514.1995.tb00533.x",

language = "English",

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journal = "Journal of Food Biochemistry",

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AU - KO, JESANG

AU - CHEAH, SHWUYENG

AU - FISCHL, ANTHONY S.

PY - 1995/8

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N2 - The membrane associated enzyme phosphatidylinositol: ceramide phosphoinositol transferase (IPC synthase) catalyzes an essential step in the biosynthesis of yeast inositol‐containing sphingolipids. A variety of solubilization agents were examined for their ability to release IPC synthase activity from yeast membranes. The most effective solubilization agent was Triton X‐100 which released over 90% of the IPC synthase activity. The basic enzymological properties of the solubilized enzyme were determined using a Triton X‐100/phosphatidylinositol/ceramide/enzyme mixed micellar assay system. IPC synthase activity was dependent upon the surface concentrations of phosphatidylinositol and ceramide in a phosphatidylinositol/ceramide/Triton X‐100 mixed micelle. Maximal enzyme activity was measured at 30C and pH 7.0 in the presence of 5 mM Triton X‐100, 1 mM manganese and 5 mM magnesium ions.

AB - The membrane associated enzyme phosphatidylinositol: ceramide phosphoinositol transferase (IPC synthase) catalyzes an essential step in the biosynthesis of yeast inositol‐containing sphingolipids. A variety of solubilization agents were examined for their ability to release IPC synthase activity from yeast membranes. The most effective solubilization agent was Triton X‐100 which released over 90% of the IPC synthase activity. The basic enzymological properties of the solubilized enzyme were determined using a Triton X‐100/phosphatidylinositol/ceramide/enzyme mixed micellar assay system. IPC synthase activity was dependent upon the surface concentrations of phosphatidylinositol and ceramide in a phosphatidylinositol/ceramide/Triton X‐100 mixed micelle. Maximal enzyme activity was measured at 30C and pH 7.0 in the presence of 5 mM Triton X‐100, 1 mM manganese and 5 mM magnesium ions.

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DO - 10.1111/j.1745-4514.1995.tb00533.x

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SN - 0145-8884

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JF - Journal of Food Biochemistry

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SOLUBILIZATION AND CHARACTERIZATION OF MICROSOMAL‐ASSOCIATED PHOSPHATIDYLINOSITOL: CERAMIDE PHOSPHOINOSITOL TRANSFERASE FROM SACCHAROMYCES CEREVISIAE

Abstract

ASJC Scopus subject areas

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